The TCO2 (total carbon dioxide) test is performed on the blood of racehorses as a means of combatting the practice of administering alkalizing agents for the purpose of enhancing performance. The purposes of this review are to present an overview of the factors contributing to TCO2 and to review the literature regarding TCO2 in adult Standardbred and Thoroughbred horses to demonstrate the range of variability of TCO2 in horses. Most of the research published on the topic of TCO2 or bicarbonate measurement in racehorses was accessed and reviewed. PubMed and Google Scholar were the primary search engines used to source the relevant literature. The main physicochemical factors that contribute to changes in TCO2 in horses at rest are changes in strong ions concentration, followed by changes in weak acid (i.e. plasma albumin) concentrations. There is a wide normal distribution of TCO2 in horses ranging from 23 mmol/L to 38 mmol/L. Independent of administration of alkalizing agents, blood TCO2 is affected mainly by feeding, time of day (diurnal variation), season and exercise. There are few studies that have reported hour-by-hour changes in TCO2. Racehorse population studies suffer from lack of validation regarding whether or not a horse was administered an alkalizing agent. It is concluded that the normal range of TCO2 in non-alkalized Standardbred and Thoroughbred horses is significantly wider than has been appreciated, that periods of elevated TCO2 appear to be normal for many horses at rest, and that a TCO2 test alone is not definitive for the purposes of determining of an alkalizing agent has been administered to a horse.